CN1414954A - Stable polymorph of N-(3-ethynylphenylamino)-6,7-bis (2-methoxyethoxy)-4-quinazolinamine hydrochloride, method of production and pharmaceutical uses thereof - Google Patents

Abstract

The present invention relates to a stable crystalline form of N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine hydrochloride designated the B polymorph, its production in essentially pure form, and its use. The invention also relates to the pharmaceutical compositions containing the stable polymorph B form of N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine as hydrochloride, as well as other forms of the compound, and to methods of treating hyperproliferative disorders, such as cancer, by administering the compound.

Description

Stable polymorph of N- (3-ethynylphenylamino) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, and preparation method and medical application thereof

This application claims priority from U.S. provisional applications 60/206,420 (filed 5/23/2000), 60/193,191 (filed 3/30/2000), 60/164,907 (filed 11/1999), the contents of which are incorporated herein by reference.

Various publications are cited in this application. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to describe in more detail the state of the art to which this invention pertains.

Background

N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine, whether in its hydrochloride or mesylate salt or in anhydrous or aqueous form, is useful for treating hyperproliferative diseases, such as cancer, in mammals.

U.S. Pat. No. 5,747,498 (approved 5/1998), the entire contents of which are incorporated herein by reference, discloses in example 20 that [6, 7-bis (2-methoxyethoxy) -quinazolin-4-yl]- (3-ethynylphenyl) amine hydrochloride is a carcinogenic and protocarcinogenic inhibitor of the erbB family of protein tyrosine kinases, such as Epidermal Growth Factor Receptor (EGFR), and is therefore useful in the treatment of hyperproliferative diseases such as cancer in humans.

The mesylate salt form thereof is described in PCT International application WO 99/55683(PCT/IB99/00612, filed 1999, 8.4), the entire contents of which are incorporated herein by reference, assigned to the common assignee, and having the structure shown below in formula 1:

it is used for the treatment of proliferative diseases and is more preferably administered parenterally, i.e. in solution, than the hydrochloride salt compound.

The mesylate compound is more soluble in aqueous compositions than the hydrochloride compound, and therefore, the mesylate compound is more easily administered parenterally. And the hydrochloride compound thereof is preferably in a solid dosage form such as a tablet and administered orally.

Summary of The Invention

The present invention relates to the polymorph and to a process for the selective preparation of the polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, particularly in the form of the stable polymorph.

The invention also relates to the novel use of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine, whether as its hydrochloride or mesylate salt, or in its anhydrous or hydrate form, as well as in its various polymorph forms, for the treatment of hyperproliferative diseases such as cancer in mammals.

Drawings

Figure 1 is an X-ray powder diffraction pattern of the hydrochloride form a polymorph (which is thermodynamically less stable) illustrating the first peak over a larger range.

Figure 2 is an X-ray powder diffraction pattern of the hydrochloride form a polymorph (which is less thermodynamically stable) which is shown in greater detail in the smaller range.

Figure 3 is an X-ray powder diffraction pattern of the hydrochloride form B polymorph (which is thermodynamically more stable) illustrating the first peak over a larger range.

Figure 4 is an X-ray powder diffraction pattern of the hydrochloride form B polymorph (which is thermodynamically more stable), which is shown in greater detail on a smaller scale.

Detailed Description

The present invention discloses a substantially homogeneous (homogeneous) crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride designated the B polymorph that exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14 and 26.91. The polymorph is further characterized by an X-ray powder diffraction pattern as shown in figure 3.

The present invention discloses a crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, designated the B polymorph, that exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14 and 26.91, which is substantially free of the polymorph designated the A polymorph. The polymorph is further characterized by an X-ray powder diffraction pattern as shown in figure 3.

The polymorph designated as the B polymorph can be in substantially pure form relative to the a polymorph.

Also disclosed are compositions containing a substantially homogeneous crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride that exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta of approximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14 and 26.91. N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride also exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta as shown in Table 3 or Table 4 below. Also, N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride in the form of the B polymorph may be characterized by the X-ray powder diffraction pattern shown in FIG. 3.

Also disclosed is a composition comprising a crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, designated the B polymorph, wherein said polymorph exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14 and 26.91, and wherein the weight percentage of the B polymorph relative to the A polymorph is at least 70%. The composition comprises at least 75% by weight, relative to the form a polymorph, of the form B polymorph; at least 80% by weight ofthe polymorph form B; at least 85% by weight of the polymorph form B; at least 90% by weight of the polymorph form B; at least 95% by weight of the polymorph form B; at least 97% by weight of the polymorph form B; at least 98% by weight of the polymorph form B; or at least 99% by weight of the polymorph form B.

The present invention further discloses a process for preparing the form B polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride by recrystallizing N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride in a solvent comprising an alcohol and water.

In the method, the recrystallization may include the steps of:

a) heating and refluxing alcohol, water and N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride to form a solution;

b) cooling the solution to about 65-70 ℃;

c) allowing the solution to settle; and

d) the form B polymorph is precipitated by further cooling the clear solution.

In this process, N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride is prepared by the following steps:

compound of formula 6

With a compound of formula 4

The compound of formula 6 is prepared by reacting the compound of formula 5 under heating in a suspension of an alkali metal and a solvent.

The preparation of the compound of formula 4 is carried out by chlorinating the compound of formula 3.

Also disclosed is a pharmaceutical composition for treating a hyperproliferative disorder in a mammal which consists essentially of a therapeutically effective amount of the polymorph form B and a pharmaceutically acceptable carrier.

The pharmaceutical composition may be suitable for oral administration. It can be made into tablet form.

Also disclosed is a method for treating a hyperproliferative disorder in a mammal comprising administering to said mammal a therapeutically effective amount of the polymorph form B.

The method can be used to treat a cancer selected from the group consisting of brain cancer, squamous cell carcinoma, bladder cancer, gastric cancer, pancreatic cancer, breast cancer, head cancer, neck cancer, esophageal cancer, prostate cancer, colorectal cancer, lung cancer, renal (renal) cancer, renal (kidney) cancer, ovarian cancer, gynecological cancer, and thyroid cancer.

The method can also be used to treat a cancer selected from the group consisting of non-small cell lung cancer (NSCLC), refractory ovarian cancer, head and neck cancer, colorectal cancer, and renal cancer.

In the method, the therapeutically effective amount may be from about 0.001 to about 100mg/kg day, or from about 1 to about 35mg/kg day.

In this method, the therapeutically effective amount may be from about 1 to about 7000 mg/day; from about 5 to about 2500 mg/day; from about 5 to about 200 mg/day; from about 25 to about 200 mg/day.

Also disclosed is a method for treating a hyperproliferative disorder in a mammal comprising administering to said mammal a therapeutically effective amount of the polymorph form B in combination with an antineoplastic agent selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, and anti-androgens.

The present invention still further discloses a method for preparing a composition comprising a substantially homogeneous crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, designated the B polymorph, that exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14 and 26.91, comprising mixing the crystalline polymorph designated as B with a carrier.

The carrier may be a pharmaceutically acceptable carrier.

Also disclosed is a process for preparing the form B polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, wherein the process comprises the step of recrystallizing N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride in a solvent comprising an alcohol.

In this method, the solvent may further contain water.

In this method, N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride is prepared by reacting a compound of formula 6

Coupling with a compound of formula 4.

In this process, the compound of formula 6 is prepared by reacting in an alkali metal and a solventBy heating the suspension of (a) to react the compound of formula 5.

In this process, the preparation of the compound of formula 4 is carried out by chlorinating the compound of formula 3.

The invention further discloses a process for preparing the polymorph form B of claim 1, comprising the steps of:

a) reacting in a solvent mixture of thionyl chloride, dichloromethane and dimethylformamide to substitute the starting quinazolinamine compound of formula 3 having a hydroxyl group chlorinated,

to provide a compound of formula 4;

b) in situ from the starting material of the compound of formula 5 by reacting the compound of formula 5 under heating in a suspension of an alkali metal and a solvent

Preparing a compound of formula 6;

c) reacting the compound of formula 6 in situ with a compound of formula 4, wherein the compound of formula 6 replaces the chlorine in the compound of formula 4 to provide N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride;

d) recrystallization of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride in alcohol gives the form B polymorph.

In this process, the chlorine substitution may be quenched in the presence of an aqueous sodium hydroxide solution, an aqueous sodium bicarbonate solution, an aqueous potassium hydroxide solution, an aqueous potassium bicarbonate solution, an aqueous potassium carbonate solution, an aqueous sodium carbonate solution, or a mixture thereof.

The present invention still further discloses a process for the preparation of the form B polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride by recrystallization, which comprises the steps of:

a) heating and refluxing alcohol, water and N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride to form a solution;

b) cooling the solution to about 65-70 ℃;

c) allowing the solution to settle; and

d) the form B polymorph is precipitated by further cooling the clear solution.

The invention further discloses a composition consisting essentially of the form a polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, wherein said form a polymorph is characterized by the following peaks in the X-ray powder diffraction pattern shown in figure 1.

Also disclosed are compositions consisting essentially of the form A polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, wherein said form A polymorph is characterized by the peaks shown in Table 1 or 2 below.

Prodrugs of any of the compounds herein are also disclosed.

The invention further discloses a method of inducing differentiation of tumor cells in a tumor, the method comprising contacting the cells with an effective amount of any one of the compounds or compositions disclosed herein.

Also disclosed is a method for treating NSCLC (non-small cell lung cancer), pediatric malignancies, cervical and other tumors caused or facilitated by Human Papillomavirus (HPV), melanoma, Barrett's esophagus (pre-malignant syndrome),adrenal and skin cancers and autoimmune diseases, neoplastic skin diseases and atherosclerosis in a mammal which comprises administering to said mammal a therapeutically effective amount of a composition comprising at least one N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine and pharmaceutically acceptable salts thereof in anhydrous and hydrated forms.

The treatment may further comprise palliative or neoadjuvant (neo-adjuvant)/adjuvant monotherapy (monotherapy); or blocking Epidermal Growth Factor Receptor (EGFR).

The method can also be used to treat EGFRvIII expressing tumors.

The method may further comprise a combination with any of chemotherapy and immunotherapy; or treating with either or both of anti-EGFR and anti-EGF antibodies simultaneously; or administering to said mammal a compound selected from the group consisting of MMP (matrix-metallo-protease), VEGFR (vascular endothelial growth factor receptor), farnesyl transferase, CTLA₄(cytotoxic T-lymphocyte antigen 4) and inhibitors of erbB2, MAb of VEGFr, rhuMAb-VEGF, erbB2MAb and avb3 Mab.

The pharmaceutical compounds used may be radiosensitizers for cancer treatment or for combination with anti-hormone therapy or for inhibiting the growth of human tumors in a radiation treatment regimen.

The invention further discloses a method for chemoprevention of basal cell carcinoma or squamous cell carcinoma in the skin of regions exposed to sunlight or for chemoprevention of a human at high risk for said carcinoma, which comprises administering to said human a therapeutically effective amount of a pharmaceutical composition comprising at least one N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine and pharmaceutically acceptable salts thereof in anhydrous and hydrate form.

Also disclosed are methods of inducing differentiation of tumor cells in a tumor, comprising contacting said cells with at least one N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine compound and anhydrous and hydrate forms of pharmaceutically acceptable salts thereof.

Accordingly, it is an object of the present invention to provide a process for preparing N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride (formula 2) in the form of its hydrochloride salt:

2 (polymorphs a and B) make them more suitable for tablet and oral administration and consist essentially of the stable polymorph (polymorph form B), and substantially pure forms of the compound in the form of the polymorph form B and the intermediate polymorph form a.

It is a further object of the present invention to provide the stable polymorph form B in the form of an orally administered pharmaceutical composition.

The stability of the hydrochloride salt should be considered for therapeutic use in patients, since differences in this regard will affect the effective dosage level and the mode of administration. It has now been found that the hydrochloride salt of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine exists in two polymorphic states, polymorph a and B. This is in contrast to the mesylate compound, which exists in three polymorphic states (mesylate polymorph A, B and C). The polymorph form B of the hydrochloride salt has been found to be the most thermodynamically stable and desirable form, and the present invention includes a polymorph form B compound in the form of a substantially pure polymorph form B, pharmaceutical compositions, particularly in tablet form, of a substantially pure polymorph form B, and alternative processes for preparing the compound.

The hydrochloride salt compound disclosed in U.S. patent 5,747,498 actually contains a mixture of the a polymorph and B, and is not preferred over the mesylate form for tablet dosage forms because of its partial reduction in stability (i.e., due to the a polymorph component).

In particular, the present invention relates to a process for the preparation of the compound N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride form, and a process for the preparation of stable form B in high yield. It has been found that the mesylate salt of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine has at least three polymorphic forms with increased stability identified as A, B and CWith different X-ray powder diffraction patterns. Hydrochloride salt form A polymorph (A)₁And A₂) And B (B)₁And B₂) The X-ray powder diffraction pattern of form (A) is shown in FIGS. 1-4 below: the spectra of fig. 1 and 3 illustrate the first peaks of a and B, respectively, to a greater extent, and the spectra of fig. 2 and 4 illustrate a and B, respectively, to a lesser extent, in greater detail.

The data in the X-ray powder diffraction patterns of FIGS. 1-4 above are set forth in tables 1-4 below:

table 1: polymorph form A

Anode: cu-wavelength 1: 1.54056 wavelength 2: 1.54439(Rel Strength: 0.500)

Range # 1-Coupled (Coupled:) 3.000-40.000 stages (StepSize): 0.040 grade

(StepTime)：1.00

Flat Width (smoothening Width): 0.300 threshold: 1.0

d(A)	I(rel)	d(A)	I(rel)	d(A)	I(rel)	d(A)	I(rel)	d(A)	I(rel)
										15.82794	100.0	6.63179	1.7	4.54453	4.8	3.61674	8.2	2.91238	3.5
14.32371	3.9	5.84901	2.1	4.19685	4.7	3.50393	9.3	2.73148	3.7
										11.74376	1.5	5.69971	2.3	4.16411	4.4	3.40200	6.0	2.60193	1.8
11.03408	1.2	5.46922	2.4	3.97273	4.7	3.35174	5.3	2.48243	1.3
										10.16026	1.4	5.21396	3.6	3.91344	12.4	3.29005	4.2	2.40227	2.2
8.98039	13.1	4.80569	3.5	3.78223	24.2	3.05178	7.1	2.31297	1.7
										7.85825	7.8	4.70077	12.2	3.67845	8.8	2.97750	3.0

Table 2: polymorph form A

Anode: cu-wavelength 1: 1.54056 wavelength 2: 1.54439(Rel Strength: 0.500)

Range # 1-coupled: 3.000-40.000 stages: 0.040 stage: 1.00

Flat width: 0.300 threshold: 1.0

2-Theta	I(rel)	2-Theta	I(rel)	2-Theta	I(rel)	2-Theta	I(rel)	2-Theta	I(rel)
										5.579	100.0	13.340	1.7	19.517	4.8	24.594	8.2	30.673	3.5
6.165	3.9	15.135	2.1	21.152	4.7	25.398	9.3	32.759	3.7
										7.522	1.5	15.534	2.3	21.320	4.4	26.173	6.0	34.440	1.8
8.006	1.2	16.193	2.4	22.360	4.7	26.572	5.3	36.154	1.3
										8.696	1.4	16.991	3.6	22.703	12.4	27.080	4.2	37.404	2.2
9.841	13.1	18.447	3.5	23.502	24.2	29.240	7.1	38.905	1.7
										11.251	7.8	18.862	12.2	24.175	8.8	30.007	3.0

Table 3: polymorph form B

Anode: cu-wavelength 1: 1.54056 wavelength 2: 1.54439(Rel Strength: 0.500)

Range # 1-coupled: 3.000-40.040 grades: 0.040 stage: 1.00

Flat width: 0.300 threshold: 1.0

d(A)	I(rel)	d(A)	I(rel)	d(A)	I(rel)	d(A)	I(rel)	d(A)	I(rel)
										14.11826	100.0	5.01567	2.5	3.86656	4.8	3.23688	0.9	2.74020	1.7
11.23947	3.2	4.87215	0.7	3.76849	2.3	3.16755	1.5	2.69265	1.7
										9.25019	3.9	4.72882	1.5	3.71927	3.0	3.11673	4.3	2.58169	1.5
7.74623	1.5	4.57666	1.0	3.63632	6.8	3.07644	1.4	2.51043	0.8
										7.08519	6.4	4.39330	14.4	3.53967	10.0	2.99596	2.1	2.47356	1.0
6.60941	9.6	4.28038	4.2	3.47448	3.7	2.95049	0.9	2.43974	0.6
										5.98828	2.1	4.20645	14.4	3.43610	3.9	2.89151	1.6	2.41068	1.1
5.63253	2.9	4.06007	4.7	3.35732	2.8	2.83992	2.2	2.38755	1.4
										5.22369	5.5	3.95667	4.5	3.31029	5.6	2.81037	2.4	2.35914	1.7

Table 4: polymorph form B

Anode: cu-wavelength 1: 1.54056 wavelength 2: 1.54439(Rel Strength: 0.500)

Range # 1-coupled: 3.000-40.040 grades: 0.040 stage: 1.00

Flat width: 0.300 threshold: 1.0

2-Theta	I(rel)	2-Theta	I(rel)	2-Theta	I(rel)	2-Theta	I(rel)	2-Theta	I(rel)
										6.255	100.0	17.668	2.5	22.982	4.8	27.534	0.9	32.652	1.7
7.860	3.2	18.193	0.7	23.589	2.3	28.148	1.5	33.245	1.7
										9.553	3.9	18.749	1.5	23.906	3.0	28.617	4.3	34.719	1.5
11.414	1.5	19.379	1.0	24.459	6.8	29.000	1.4	35.737	0.8
										12.483	6.4	20.196	14.4	25.138	10.0	29.797	2.1	36.288	1.0
13.385	9.6	20.734	4.2	25.617	3.7	30.267	0.9	36.809	0.6
										14.781	2.1	21.103	14.4	25.908	3.9	30.900	1.6	37.269	1.1
15.720	2.9	21.873	4.7	26.527	2.8	31.475	2.2	37.643	1.4
										16.959	5.5	22.452	4.5	26.911	5.6	31.815	2.4	38.114	1.7

It will be appreciated that the X-ray powder diffraction pattern is but one of many ways to identify the atomic arrangement of the compound comprising N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, and that other methods known in the art, such as single crystal X-ray diffraction, may be used to identify samples, compositions or other articles in which the B-form polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride is present.

The present invention relates to a compound of the form B polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride exhibiting an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14 and 26.91. The invention also relates to the polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride which exhibits an X-ray powder diffraction pattern having characteristic peaks at 2-theta degrees approximately as expressed in the values of Table 4 above.

The present invention is directed to a compound of the form a polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride that exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at approximately 5.58, 9.84, 11.25, 18.86, 22.70, 23.50, 24.18, 24.59, 25.40 and 29.24. The invention also relates to the polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride which exhibits an X-ray powder diffraction pattern having characteristic peaks at 2-theta degrees approximately as expressed in the values of Table 2 above. Preparation method

According to the process of the present invention, the substantially pure polymorph form B of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride (the compound of formula 1) is prepared by the following steps:

1) as a substituted chlorinated quinazoline amine compound having a hydroxyl group (formula 3) by performing a reaction in a solvent mixture of thionyl chloride, dichloromethane and dimethylformamide:

finally the reaction is quenched with aqueous sodium hydroxide or sodium bicarbonate. Substitution of chlorine for hydroxyl in high yieldPreparing a compound of formula 4;

2) starting from a compound of formula 5 by reacting the compound of formula 5 under heating in a suspension of NaOH (or KOH, or a combination thereof) in toluene

Preparing a compound of formula 6;

3) reacting the compound of formula 6 with the compound of formula 4 in step 1, wherein the compound of formula 6 replaces chlorine in the compound of formula 4, to give N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride (the compound of formula 2) in 97% yield;

4) recrystallization of the compound of formula 2 (containing the a form polymorph and the B form polymorph) in a solvent containing an alcohol (e.g., 2B-ethanol) and water generally yields a more stable B formpolymorph in high yield, e.g., about 85%.

Accordingly, the present invention relates to a process for the preparation of the form B polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride which comprises recrystallising N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) 4-quinazolinamine hydrochloride in a solvent comprising alcohol and water. In one embodiment, the method comprises the steps of: heating and refluxing alcohol, water and N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride to form a solution; cooling the solution to about 65-70 ℃; allowing the solution to settle; the form B polymorph is precipitated by further cooling the clear solution. In one embodiment, the alcohol is ethanol. In a preferred embodiment, the ratio of ethanol to water is 4: 1. Other lower alcohols such as C are contemplated₁-C₄Alcohols are also suitable for recrystallization of the form B polymorph, with the ratio of alcohol to water being adjusted as desired. In anotherIn a preferred embodiment, the compound to be recrystallized is present in an amount of about 0.05 (weight/volume) of the total volume of the solvent. In one embodiment, N- (3-ethynylbenzene is prepared by coupling a compound of formula 6 with a compound of formula 4Yl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride. In another embodiment, the compound of formula 6 is prepared by reacting the compound of formula 5 under heating in a suspension of an alkali metal and a solvent.

In one embodiment, the compound of formula 4 is prepared by reacting the compound of formula 3 in a solvent mixture of thionyl chloride, dichloromethane and dimethylformamide to chlorinate it, followed by quenching the reaction with aqueous sodium hydroxide. Or an aqueous sodium bicarbonate solution may be substituted for the sodium hydroxide solution.

The present invention relates to the N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride polymorph form B prepared by the above method. In one embodiment, the polymorph form B is prepared from the starting materials described herein. In a preferred embodiment, the polymorph form B is prepared by reacting the starting materials described herein with the reagents and conditions described in the methods described herein and in the examples that follow. General Synthesis

It has been found that N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride exists in two different anhydrous polymorphic forms, form a and B. The preparation of the various polymorphs is carried out by reacting the ingredients separately according to the following schemes.Applications of

As described in the above-mentioned U.S. patent No. 5,747,498 and PCT WO 99/55683, the compounds according to the present invention may be used for the treatment of proliferative diseases in mammals, comprising a therapeutically effective amount of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride or mesylate and a pharmaceutically acceptable carrier.

The term "compound of the invention" as used herein is preferably the form B polymorph of the hydrochloride salt N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, but is not meant to exclude the mesylate salt form or its three polymorphs, or its hydrochloride form a polymorph, or a mixture of its hydrochloride forms B and a polymorph, or other non-crystalline forms of the compound.

The term "treating" as used herein, unless otherwise indicated, means reversing, alleviating, inhibiting the disease or disorder to which the term applies, or preventing the disease or disorder, or one or more symptoms of the disease or disorder. The term "treatment" as used herein refers to the action of treatment (treating), wherein the treatment is as defined herein before.

As used herein, "abnormal cell growth" refers to growth that is independent of normal cellular regulatory mechanisms (e.g., loss of contact inhibition), which includes abnormal growth of normal cells and growth of abnormal cells. It includes, but is not limited to, the following abnormal growths: (1) tumor cells (tumors), including benign and malignant, expressing the active Ras tumor gene; (2) tumor cells (tumors), both benign and malignant, in which the Ras protein is activated due to an oncogenic mutation of another gene; (3) other proliferative diseases of benign and malignant cells, in which abnormal Ras activation occurs. Examples of such benign proliferative diseases are psoriasis, benign prostatic hypertrophy, Human Papilloma Virus (HPV) and restenosis. As used herein, "abnormal cell growth" also includes abnormal growth of cells, including benign and malignant, due to activation of the following enzymatic enzymes, nisin transferase, protein kinase, protein phosphatase, lipid kinase, lipid phosphatase, or transcription factors, or activation of intracellular or cell surface receptor proteins.

[6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride, preferably a stable B-type polymorph, and additionally for the treatment of a variety of other human tumors containing hyperproliferative cells activated by EGFR-stimulated signaling pathways, whether EGFR protein itself is overexpressed (e.g., due to one or more altered transcription, altered mRNA degradation, or gene expansion), using another receptor protein EGFR, a ligand that activates EGFR (e.g., EGF, TGF α, amphiregulin, β -animal cellulose, heparin-bound EGF, or epiregulin, or heterodimeric receptor, or is dependent or partially dependent on EGFR protein "normal" level activity, whether activation is by extracellular ligand, intracellular signaling pathway and/or genetic alteration, or a polymorph that results in increased or ligand-dependent activity of anal substitutions (e.g., RvIII, Clin, and/or genetic alterations), including renal cell lymphoma, esophageal adenocarcinoma, pancreatic adenocarcinoma, renal cell lymphoma, pancreatic adenocarcinoma, prostate adenocarcinoma, bladder adenocarcinoma, prostate adenocarcinoma.

[6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride, preferably the stable polymorph form B, is additionally useful in the treatment of a variety of other human proliferative disorders in mammals which contain hyperproliferative cells activated by signal transduction pathways capable of being stimulated by EGFR, such as benign hyperplasia of the skin (e.g., psoriasis) or prostate (e.g., BPH), chronic pancreatitis, or hyperplasia of pancreatic duct epithelial cells, renal disorders (including proliferative glomerulonephritis and diabetes-induced renal disorders), wherein the composition contains a therapeutically effective amount of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, preferably the polymorph form B, and a pharmaceutically acceptable carrier.

In addition, a composition comprising a compound prepared according to the present invention, wherein the composition comprises a therapeutically effective amount of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, preferably the polymorph form B, and a pharmaceutically acceptable carrier, provides prophylaxis of blast transplantation in mammals.

[6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride, preferably a stable B-type polymorph, is additionally useful for the treatment of other diseases in which cells are activated by EGFR-stimulated signal transduction pathways, whether by overexpression of EGFR protein itself (e.g., due to one or more altered transcription, altered mRNA degradation, or gene expansion), which can form heterodimers using another receptor protein, or which activate a ligand for EGFR (e.g., EGF, TGF α, amphiregulin, β -animal cellulose, heparin-bound EGF, or epiregulin), or which is dependent or partially dependent on the "normal" level of EGFR protein, whether activation is by extracellular ligands, intracellular signal transduction pathways, and/or genetic alterations, or which is a polymorph, which results in amino acid substitutions that produce increased or ligand-dependent activity (e.g., EGFR RvIII, Archer G.E. 1999) Clinical pathways and/or genetic alterations, or which results in the proliferation of cells involved in the proliferation of epithelial cells which are involved in the regeneration of epithelial cells, proliferation of epithelial cells which are involved in the proliferation of epithelial cell proliferation, cell proliferation of epithelial cell proliferation, cell proliferation of epithelial cell proliferation, cell proliferation of epithelial cell type III, tumor cell proliferation of lung, tumor cell proliferation of epithelial cell type III, tumor cell proliferation of lung, tumor cell proliferation of lung tumor, tumor cell proliferation of lung tumor, tumor cell proliferation of lung tumor, kidney tumor cell proliferation of lung tumor, and lung tumor, tumor cell proliferation of lung.

[6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride, preferably the stable polymorph form B, is additionally useful for the treatment of various leukemias (chronic and acute) and lymphoid malignancies (e.g., lymphocytic lymphoma), diabetes and other retinopathies, such as retinophay or prematurity, age-related macular degeneration, childhood solid tumors, gliomas, hemangiomas, melanomas, including intraocular or uveal melanoma, Kapos's sarcoma, Hodgkin's disease, epidermoid carcinoma, cancers of the endocrine system (e.g., parathyroid carcinoma, adrenal gland carcinoma), small bone (bonemall intestine) carcinoma, urinary tract carcinoma, penile carcinoma, and ureter carcinoma, atherosclerosis, skin diseases such as eczema and scleroderma, mycoses (mycoses) fungoides, soft tissue sarcomas, and central nervous system tumors (e.g., primary CNS lymphoma, spinal tumors, brain stem glioma, or pituitary adenoma).

The treatment of any of the above hyperproliferative or other diseases may employ monotherapy of [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride, preferably the stable form B polymorph thereof, and may involve one or more other drugs or treatments (e.g. radiation therapy, chemotherapy) in addition to it which are anti-hyperproliferative, anti-tumour or anti-proliferative in nature. Such combination therapy may be achieved by the simultaneous, sequential, cyclic or divided dose administration of the therapeutic homogeneous ingredients. For any of the above treatments, [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride, preferably the stable form B polymorph thereof, is typically used at a dose of 1-7000 mg/day, preferably 5-2500 mg/day, most preferably 5-200 mg/day.

Also, various types of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamines, including the mesylate and hydrochloride forms (all polymorphs) as well as other pharmaceutically acceptable salt forms and anhydrate and hydrate forms, can be used to treat specific disorders of NSCLC (non-small cell lung cancer), pediatric malignancies, cervical cancer, and other tumors caused or facilitated by Human Papilloma Virus (HPV), melanoma, Barrett's esophageal tumor (pre-malignant syndrome), adrenal and skin cancers, and autoimmune and skin tumor diseases such as mold fungi, as well as basic chemopreventionor cutaneous squamous cell carcinoma, particularly in areas exposed to sunlight or in persons known to be at high risk for, e.g., cancer. In addition, the above compounds are useful in the treatment of atherosclerosis, which has an epidermal growth factor implicated in the hyperproliferation of vascular smooth muscle cells that can cause atherosclerotic plaques (G.E. Peoples et al, Proc. Nat. Acad. Sci. USA 92: 6547-.

The compounds of the present invention are potent inhibitors of the erbB family of oncogenic and proto-oncogenic (protooncogenic) protein tyrosine kinases, such as Epidermal Growth Factor Receptor (EGFR), erbB2, HER3, or HER4, and are therefore suitable for therapeutic use as antiproliferative (e.g. anticancer) agents in mammals, particularly humans. The compounds of the present invention are also inhibitors of angiogenesis and/or vasculogenesis.

The compounds of the present invention may also be useful in the treatment of a variety of other diseases in which a variety of aberrantly expressed ligand/receptor interactions or activation or signaling events associated with a variety of protein tyrosine kinases are implicated. These diseases may include diseases involving aberrant function, expression, activation or signalling of erbB tyrosine kinase neurons, glia, astrocytes, hypothalamus, glands, macrophages, epithelium, stroma or blastocoel properties. In addition, the compounds of the invention may have therapeutic utility in inflammatory, angiogenic and immune diseases involving identified and unidentified tyrosine kinases that are capable of being inhibited by the compounds of the invention.

These generalities are in addition to the direct treatment of the above-mentioned diseases with the compoundsCan be used as a palliative or neoadjuvant/adjuvant monotherapy in blocking Epidermal Growth Factor Receptor (EGFR), for the treatment of tumors expressing EGFR variants knownas EGFRvIII (e.g., DKMOSCATELO et al, Cancer Res. 55: 5536-5539, 1995), and in combination with chemotherapy and immunotherapy. As described in more detail below, the treatment may also be combined with anti-EGFR and anti-EGF antibodies, or with inhibitors of: MMP (matrix-metallo-protease), other tyrosine kinases including VEGFR (vascular endothelial growth factor receptor), farnesyl transferase, CTLA₄(cytotoxic T-lymphocyte antigen 4), and erbB 2. Further treatments include mabs to VEGFr, and other cancer-associated antibodies, which include rhuMAb-VEGF (Genentech, stage III), erbB2Mab under the trade name Herceptin (Genentech, stage III), or avb3 Mab under the trade name Vitaxin (Applied Molecular Evolution/medimmunee, stage II).

The invention is also directed to a pharmaceutical composition and method of treating any of the foregoing in a mammal comprising administering to said mammal a therapeutically effective amount of the form B polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine, preferably the hydrochloride salt, and a pharmaceutically acceptable carrier. Combination therapy

The active compounds may be used as monotherapy or may be combined with one or more substances or therapeutic agents such as anti-EGFR and anti-EGF antibodies, or with inhibitors of: MMP (matrix-metallo-protease), other tyrosine kinases including VEGFR (vascular endothelial growth factor receptor), farnesyl transferase, CTLA₄(cytotoxic T-lymphocyte antigen 4), and erbB2, as well as mabs to VEGFr, and other related antibodies, including rhuMAb-VEGF, erbB2Mab, or avb 3.

Thus, the active compound may be used together with one or more other anti-neoplastic agents, for example selected from, for example, mitotic inhibitors, such as vinblastine; alkylating agents such as cisplatin, carboplatin, and cyclophosphamide; antimetabolites, for example 5-fluorouracil, cytarabine and hydroxyurea, or one of the preferred antimetabolites, for example as disclosed in European patent application 239362, such as N- (5- [ N- (3, 4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino]-2-thenoyl) -L-glutamic acid; a growth factor inhibitor; a cell cycle inhibitor; insertion antibiotics, such as doxorubicin and bleomycin; enzymes, such as interferon; and anti-hormonal agents, e.g. anti-estrogens such as tamoxifen (D (tamoxifen), or e.g. anti-androgens such as carvedilol (4 '-cyano-3- (4-fluorophenylsulphonyl) -2-hydroxy-2-methyl-3' - (trifluoromethyl) propionylaniline).

In a further embodiment, in the methods of treatment with the compositions described herein, the compounds of the present invention may be used in combination with one or more anti-angiogenic agents, such as one or more MMP-2 (matrix-metallo-protease-2) inhibitors, one or more MMP-9 (matrix-metallo-protease-9) inhibitors, and/or one or more COX-II (cyclooxygenase II) inhibitors. For combination therapy with the compositions of the present invention, one of ordinary skill in the art can determine the effective amount of a compound of the present invention and chemotherapy or other agents used to inhibit abnormal cell growth (e.g., other antiproliferative agents, antiangiogenic agents, signal transduction inhibitors, or immune system enhancing agents) based on the effective amounts of the compounds described herein and those known or described compounds. The dosage form and route of administration of these therapeutic agents and compositions can be determined based on the information described for the compositions and medicaments containing the compounds of the present invention as the sole active ingredient and the information provided for the chemotherapeutic or other agents combined therewith.

The present invention also relates to the production of compounds for use in a method of treating a hyperproliferative disease in a mammal comprising administering to said mammal a therapeutically effective amount of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride in combination with an anti-neoplastic agent selected from the group consisting of: mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, and anti-androgens.

The compounds may also be used as radiosensitizers for cancer therapy and may be combined with anti-hormone therapy. Parameters for adjuvant radiation therapy are disclosed, for example, in PCT/US99/10741, published 25.11.1999, International publication No. WO 99/60023, the disclosure of which is incorporated herein by reference. For this type of treatment, for example for the purpose of inhibiting tumor growth, a radiation dose of 1-100 Gy is used, preferably in combination with at least 50mg of the pharmaceutical compound, with a preferred dosage regimen of at least 5 days per week for 2-10 weeks. Thus, the present invention further relates to a method for inhibiting abnormal cell growth in a mammal, which comprises administering to the mammal an amount of a compound of the present invention, or a pharmaceutically acceptable salt, solvate or prodrug thereof, in combination with radiation therapy, wherein the amount of the compound, salt, solvate or prodrug in combination with the radiation therapy is effective to inhibit abnormal cell growth in the mammal. Techniques for the administration of radiation therapy are well known to those of ordinary skill in the art and may be used for the combination therapies described herein.

In the methods and pharmaceutical compositions described herein, anti-angiogenic agents such as MMP-2 (matrix-metallo-protease-2) inhibitors, MMP-9 (matrix-metallo-protease-9) inhibitors, and COX-II (cyclooxygenase II) inhibitors may be used in combination with the compounds of the present invention. Examples of useful COX-II inhibitors include CELEBREXM (alecoxib), valdecoxib, and rofecoxib. Examples of useful matrix-metallo-protease inhibitors are disclosed in the following patents: WO 96/33172 (published 24/10/1996), WO 96/27583 (published 7/3/1996), European patent application No.97304971.1 (published 8/7/1997), European patent application No.99308617.2 (published 29/10/1999), WO 98/07697 (published 26/2/1998), WO98/03516 (published 29/1/1998), WO 98/34918 (published 13/8/1998), WO 98/34915 (published 13/8/1998), WO 98/33768 (published 6/8/1998), WO 98/30566 (published 16/7/1998), European patent application 606,046 (published 13/7/1994), European patent application 931,788 (published 28/7/1999), WO90/05719 (published 331/1990), WO 99/52910 (published 21/10/1999), WO 99/52889 (published 21/1999), WO 99/29667 (published 17.6.1999), PCT International application No. PCT/IB98/01113 (filed 21.7.1998), European patent application No.99302232.1 (filed 25.3.1999), British patent application No. 9912961.1 (filed 3.3.1999), U.S. provisional application No. 60/148,464 (filed 12.8.1999), U.S. patent 5,863,949 (approved 26.26.1.1999), U.S. patent 5,861,510 (granted 19.1.19.1999), and European patent publication No. 780,386 (published 25.6.1997), all of which are incorporated herein by reference. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity for inhibiting MMP-1. More preferred are those that selectively inhibit MMP-2 and/or MMP-9 relative to other matrix-metallo-proteases (i.e., MMP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).

Some specific examples of MMP inhibitors useful in the present invention are AG-3340, RO 32-3555, RS 13-0830, and the compounds listed below:

3- [ [4- (4-fluoro-phenoxy) -benzenesulfonyl]- (1-hydroxycarbamoyl-cyclopentyl) -amino]-propionic acid;

3-exo-3- [4- (4-fluoro-phenoxy) -benzenesulfonylamino]-8-oxa-bicyclo [3.2.1]octane-3-carboxylic acid hydroxyamide;

(2R, 3R)1- [4- (2-chloro-4-fluoro-benzyloxy) -benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide;

4- [4- (4-fluoro-phenoxy) -benzenesulfonylamino]-tetrahydropyran-4-carboxylic acid hydroxyamide;

3- [ [4- (4-fluoro-phenoxy) -benzenesulfonyl]- (1-hydroxycarbamoyl-cyclobutyl) -amino]-propionic acid;

4- [4- (4-chloro-phenoxy) -benzenesulfonylamino]-tetrahydropyran-4-carboxylic acid hydroxyamide;

(R)3- [4- (4-chloro-phenoxy) -benzenesulfonylamino]-tetrahydropyran-3-carboxylic acid hydroxyamide;

(2R, 3R)1- [4- (4-fluoro-2-methyl-benzyloxy) -benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide;

3- [ [4- (4-fluoro-phenoxy) -benzenesulfonyl]- (1-hydroxycarbamoyl-1-methyl-ethyl) -amino]-propionic acid;

3- [ [4- (4-fluoro-phenoxy) -benzenesulfonyl]- (4-hydroxycarbamoyl-tetrahydropyran-4-yl) -amino]-propionic acid;

3-exo-3- [4- (4-chloro-phenoxy) -benzenesulfonylamino]-8-oxa-bicyclo [3.2.1]octane-3-carboxylic acid hydroxyamide;

3-endo-3- [4- (4-fluoro-phenoxy) -benzenesulfonylamino]-8-oxa-bicyclo [3.2.1]octane-3-carboxylic acid hydroxyamide; and

(R)3- [4- (4-fluoro-phenoxy) -benzenesulfonylamino]-tetrahydrofuran-3-carboxylic acid hydroxyamide;

and pharmaceutically acceptable salts and solvates of said compounds.

Other anti-angiogenic agents, including other COX-II inhibitors and other MMP inhibitors, may also be used in the present invention.

The compounds of the invention may also be used with signal transduction inhibitors, such as drugs that inhibit the EGFR (epidermal growth factor receptor) response, such as EGFR antibodies, EGF antibodies, and other molecules that are EGFR inhibitors; VEGF (vascular endothelial growth factor) inhibitors, such as VEGF receptors and molecules that inhibit VEGF; and erbB2 receptor inhibitors, e.g. other organic molecules or antibodies that bind to the erbB2 receptor, e.g. HERCEPTIN^TM(Genentech，Inc.of South SanFrancisco，California，USA)。

EGFR inhibitors are disclosed, for example, in the following documents: WO95/19970 (published 27/7/1995), WO 98/14451 (published 9/4/1998), WO 98/02434 (published 22/1/1998), and other compounds described in U.S. Pat. No. 5,747,498 (granted 5/1998), which may be used in the invention described herein. EGFR inhibitors include, but are not limited to, monoclonal antibody C225 and anti-EGFR 22Mab (Imclone systems incorporated of New York, New York, USA), compound ZD-1839(AstraZeneca), BIBX-1382(Boehringer Ingelheim), MDX-447 (Mediarex of Annalale, New Jersey, USA), and OLX-103(Merck&Co. of Whitehouse State, New Jersey, USA), VRCTC-310(Ventech Research) and EGF fusion toxins (Seragen incorporated of Hopkinton, Massachusetts). These and other EGFR inhibitors may be used in the present invention.

VEGF inhibitors, such as SU-5416 and SU-6668(Sugen Inc. of South san Francisco, Calif., USA) may also be combined with the compounds of the present invention. VEGF inhibitors are disclosed, for example, in the following documents: for example, WO 99/24440 (published 20/5/1999), PCT International application PCT/IB99/00797 (filed 3/5/1999), WO 95/21613 (published 17/8/1995), WO 99/61422 (published 2/12/1999), US patent 5,834,504 (granted 10/11/1998), WO 98/50356 (published 12/11/1999), US patent 5,883,113 (granted 16/1999), US patent 5,886,020 (granted 23/3/1999), US patent 5,792,783 (granted 11/1998), WO99/10349 (published 4/1999), WO 97/32856 (published 12/1997), WO 97/22596 (published 26/1997 6/1998), WO 98/54093 (published 3/1998), WO 98/02438 (published 22/1/1998), WO 99/16755 (published 4/1999), And WO 98/02437 (published on 22/1/1998), all of which are incorporated herein by reference. Other examples of some specific VEGF inhibitors for use in the present invention are IM862(Cytran inc. of Kirkland, Washington, USA); genentech's anti-VEGF monoclonal antibody, inc. of South San Francisco, California; and angiozyme, a synthetic ribozyme from a ribozyme source (Boulder, Colorado) and Chiron (Emeryville, California). These and other EGFR inhibitors may be used in the present invention.

ErbB2 receptor inhibitors, such as GW-282974(Glaxo Wellcome plc) and monoclonal antibodies AR-209(Aronex Pharmaceuticals Inc. of The Woodlands, Texas, USA) and 2B-1(Chiron) can be further combined with The compounds of The present invention, such as those disclosed in: WO 98/02434 (published 22/1/1998), WO 99/35146 (published 15/7/1999), WO 99/35132 (published 15/7/1999), WO 98/02437 (published 22/1/1998), WO 97/13760 (published 17/4/1997), WO95/19970 (published 27/7/1998), U.S. Pat. No. 5,587,458 (granted 24/12/1996), U.S. Pat. No. 5,877,305 (granted 2/3/1999), the entire contents of which are incorporated herein by reference. ErbB2 receptor inhibitors useful in the present invention are also disclosed in U.S. provisional application 60/117,341 (filed 1/27/1999) and U.S. provisional application 60/117,346 (filed 1/27/1999), both of which are incorporated herein by reference. In accordance with the present invention, the erbB2 receptor inhibitor compounds and substances described in the above-mentioned PCT applications, U.S. patents, and U.S. provisional applications, as well as other compounds and substances capable of inhibiting the erbB2 receptor, may be used in combination with the compounds of the present invention.

The compounds of the present invention may be used in combination with other drugs for the treatment of abnormal cell growth or cancer, including but not limited to: drugs capable of enhancing anti-tumor immune responses, such as CTLA4 (cytotoxic lymphocyte antigen 4) antibodies and other drugs capable of blocking CTLA 4; and antiproliferative agents such as farnesyl protein transferase inhibitors. Specific CTLA4 antibodies that can be used in the present invention include those described in us provisional application 60/113,647 (filed 12/23 of 1998), which is incorporated herein by reference in its entirety, although other CTLA4 antibodies can also be used in the present invention.

These combinations may be achieved by treatment with the individual compounds at simultaneous, sequential or separate doses.

It is contemplated that the compounds of the present invention may render abnormal cells more susceptible to radiation therapy to better kill and/or inhibit the growth of these cells. Accordingly, the present invention further relates to a method of sensitizing abnormal cells in radiation therapy in a mammal comprising administering to the mammal an amount of a compound of the present invention, a pharmaceutically acceptable salt or solvate or prodrug thereof, wherein the amount is sufficient to sensitize the abnormal cells to the radiation therapy. The dosage of the compounds, salts or solvates or prodrugs thereof described in this method can be determined according to the method described herein for determining an effective dosage of the compounds of the invention.

The subject of the invention is also isotopically-labelled compounds, which are identical to those recited above for the present invention, but for the fact that one or more atoms are replaced by an atom having an atomic mass and mass number different from the atomic mass and mass number usually found in practice. Examples of isotopes that can be incorporated into compounds of the invention include hydrogen, carbon, nitrogen and oxygen, respectivelyIsotopes, e.g. of²H、³H、¹³C、¹⁴C、¹⁵N、¹⁸O and¹⁷and O. Compounds of the invention containing the aforementioned isotopes and/or other isotopes of other atoms, as well as pharmaceutically acceptable salts of such compounds, are also within the scope of the invention. Certain isotopically-labelled compounds of the present invention, e.g. radioactive isotopes¹³C and¹⁴c incorporated thereinThese may be used for drug and/or substrate tissue distribution detection. Tritium (i.e. tritium³H) And¹⁴c is particularly preferred because of its ease of preparation and detection. And, by heavier isotopes such as deuterium (i.e. by deuterium)²H) Instead, certain therapeutic advantages may be obtained due to higher metabolic stability, such as increased half-life or reduced dosage requirements, and thus may be preferred in some instances. Isotopically-labeled compounds of the present invention can be prepared by employing the procedures of the present invention and/or the procedures described in the examples below, except that readily available isotopically-labeled reagents are substituted for non-isotopically-labeled reagents using procedures well known in the art. Accordingly, the compounds of the invention for use in the methods of treatment and pharmaceutical compositions described herein also include isotopically-labeled forms of the compounds.

[6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride, preferably the stable polymorph form B, is typically used in a dose of 1-7000 mg/day, preferably 5-2500 mg/day, more preferably 5-200 mg/day in any of the above treatments.

Patients that may be treated with the compounds of the invention, alone or in combination, include, for example, patients diagnosed with: psoriasis, BPH, lung, bone, pancreatic, skin, head and neck, skin or intraocular melanoma, ovarian, rectal, anal region, stomach, colon, breast, gynecological (e.g., uterine sarcoma, fallopian tube, endometrial, cervical, vaginal, or vulval), Hodgkin's disease, esophageal, small intestine, endocrine (e.g., thyroid, parathyroid, or adrenal), soft tissue sarcoma, urinary tract, penile, prostate, chronic or acute leukemia, solid tumor in childhood, lymphocytic lymphomas, bladder, kidney or ureter (e.g., renal cell carcinoma, renal pelvis), or central nervous system (e.g., primary CNS lymphoma, spinal axis, brain stem glioma, or pituitary adenoma. active melanoma)

The in vitro activity of the compounds of the invention in inhibiting receptor tyrosine kinases (and thus subsequent proliferative diseases) can be determined by the following methods:

in vitro activity of the compounds of the invention was determined by measuring the phosphorylation inhibition of exogenous substrates of tyrosine (e.g., Lys 3-gateway or polyGluTyr (4: 1) random copolymer (I.Posner et al, J.biol.chem.267(29), 20638-47(1992)) by epidermal growth factor receptor kinase in comparison to controls affinity purified, soluble human EGF receptor (96 ng) was prepared from A431(American Type Culture Collection, Rockville, Md.) by the method of G.N.Gill, W.Weber in Enzymoloay 146, 82-88(1987)In the presence of phosphate buffer + vanadate (PBV: 50mM HEPES, pH 7.4; 125mM NaCl; 24mM MgCl)₂(ii) a 100 μ M sodium orthovanadate) in a total volume of 10 μ l at room temperature for 20-30 min. Test compounds dissolved in dimethyl sulfoxide (DMSO) were diluted with PBV, 10. mu.l were mixed with EGF receptor/EGF mixture and incubated at 30 ℃ for 10-30 minutes. By adding 20. mu.l to the EGFr/EGF mixture³³P-ATP/substrate mixture (120. mu.M Lys3-Gastrin (one letter sequence numberof amino acid KKKGPWLEEEEEAYGWLDF), 50mM Hepes pH7.4, 40. mu.M ATP, 2. mu. Ci. gamma. -, 2³³P]ATP) and incubation at room temperature for 20 min. The reaction was stopped by adding 10. mu.l of a stop solution (0.5M EDTA, pH 8; 2mM ATP) and 6. mu.l of 2N HCl. The tube was centrifuged at 14,000 RPM for 10 minutes at 4 ℃. 35 μ L of supernatant was pipetted from each tube onto a 2.5cm circle of Whatman P81 paper, washed four times with 1L each in 5% acetic acid and dried. This results in binding of the substrate to the paper due to removal of ATP in the wash. Determination of bound by liquid scintillation counting³³P]. The relative values were calculated by subtracting the binding value without substrate (e.g., lys3-gastrin) from all values as backgroundPercent inhibition over control (no test compound). These assays, performed over a range of test compound doses, can determine the approximate IC of EGFR kinase activity inhibition in vitro₅₀。

Other methods for determining the activity of the compounds of the present invention are described in U.S. Pat. No. 5,747,498, the disclosure of which is incorporated herein by reference. Pharmaceutical composition

The pharmaceutical compositions are most preferably in a form suitable for oral administration as tablets, capsules, pills, powders, sustained release dosage forms, solutions and suspensions, for example. The compositions are less preferred (the mesylate salt is preferred form) in the form of parenteral injections as sterile solutions, suspensions or emulsions, topical administration as ointments or emulsions, rectal administration as suppositories. The pharmaceutical composition may be in unit dosage form suitable for single dose administration of precise dosages. The pharmaceutical compositions will include conventional pharmaceutically acceptable carriers or excipients and the compounds according to the invention as active ingredients. In addition, it may include other drugs or agents, carriers, adjuvants, etc.

Parenteral administration forms which may be practiced include solutions or suspensions of the active compound in sterile aqueous solution, for example, propylene glycol aqueous solution or dextrose solution. The dosage form may be suitably buffered if desired.

Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents. The pharmaceutical composition may contain other ingredients such as flavoring agents, binders, excipients, and the like, if necessary. Thus, for oral administration, tablets may be employed which contain various excipients such as citric acid, disintegrating agents such as starch, alginic acid and certain complex silicates, and binding agents such as sucrose, gelatin and acacia. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate and talc are also commonly used in tablets. Solid compositions of similar form may also be used with soft and hard filled gelatin capsules. Thus, preferred materials include lactose or milk sugar and high molecular weight polyethylene glycols. When aqueous suspensions or elixirs are desired for oral administration of the compounds of the invention, they may contain various sweetening agents, flavoring agents, coloring agents or dyes, and if desired emulsifying agents or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin or combinations thereof. In addition, the compounds of the present invention may be administered topically and may be formulated as creams, gels, ointments, salves, etc. in accordance with standard pharmaceutical practice.

The compounds of the present invention may also be administered to mammals other than humans. The dosage to be administered to the mammal is determined according to the species of animal or the disease or condition to be treated. The compounds can be administered to the animal in the form of capsules, boluses, tablets, or liquid drenches. The compounds can also be administered to animals by injection or implantation. These dosage forms are prepared according to standard veterinary procedures. Alternatively, the compound may be administered with animal feed, for which purpose a feed additive or additive premix may be prepared for mixing with conventional animal feed.

Methods for preparing a variety of pharmaceutical compositions containing specific amounts of active compounds will be readily apparent to those of ordinary skill in the art. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easter, Pa., 15th Edition (1975). Administration and dosage

The compounds of the present invention (hereinafter "active compounds") can be effectively administered by any method that is capable of delivering the compound to the active site. These methods preferably include oral routes of administration, such as in tablets, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion solutions), topical and rectal administration. The hydrochloride salt form B polymorph is preferably administered orally, although parenteral administration is generally preferred.

The amount of active compound administered will be determined by the patient being treated, the severity of the disease or condition, the rate of administration and the judgment of the prescribing physician. However, for single or divided doses, the effective dosage range is about 0.001-100mg/kg body weight/day, preferably about 1-35mg/kg day. For a 70kg human, the dosage is about 0.05-7 g/day, preferably about 0.2-2.5 g/day. In some instances, dosage levels below the lower limit of the aforesaid range are effective, while in other instances, higher doses may be employed without adverse effects,provided that the higher dose is initially divided into several small doses for administration throughout the day.

[6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride, preferably the stable form B polymorph, may be used to treat patients (experimentally determined, e.g. to increase survival time) by combination therapy at a dose of 1-7000 mg/day, preferably 5-2500 mg/day, most preferably 5-200 mg/day, e.g. NSCLC (IIIb/V) with carboplatin/paclitaxel or gemcitabine/cisplatin as first line (1)^stline), NSCLC (IIIb/V) as a second line treatment for head and neck cancer with taxotere, and methotrexate as a second line treatment for patients with 5 FU/cisplatin that are refractory to control.

[6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride, preferably the stable form B polymorph, at a dose of 1-7000 mg/day, preferably 5-2500 mg/day, most preferably 5-200 mg/day may also be used for the treatment of other conditions, including pancreatic cancer, with or without combination therapy with gemcitabine, as first line treatment for the following diseases: renal cancer, gastric cancer, prostate cancer, colorectal cancer (e.g., as second line therapy for patients who fail 5FU/LCV/Irinotecan therapy), hepatocellular cancer, bladder cancer, brain cancer, ovarian cancer, breast cancer, and cervical cancer. For these treatments, patients with disease whose condition is difficult to control can be readily treated effectively by increasing response time, increasing time to progression or increasing survival time.

For any of the above treatments, [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride, the preferably stable form B polymorph, is typically used at a dose of 1-7000 mg/day, preferably 5-2500 mg/day, most preferably 5-200 mg/day.

The examples and preparations provided belowfurther illustrate the compounds of the present invention and methods of preparing these compounds. The present invention can be better understood by the following experiments thereof. It will be apparent to those of ordinary skill in the art that these specific methods and results of the discussion are merely illustrative of the invention, which is defined in more detail by the claims that follow.

Embodiment example 1: preparation of the Compound of formula 4

The following materials were used in the synthesis of compounds of formula 4:

material	Measurement of	Unit of	Equivalent weight/volume
				A compound of formula 3	88.0	kg	1 equivalent of
Thionyl chloride	89.0	kg	2.5 equivalents
				Dimethyl formamide
	11	kg	0.5 equivalent
				Methylene dichloride	880.0	L	10L/kg
50% sodium hydroxide aqueous solution	According to the need	L						1 equivalent of
				Heptane (Heptane)	880.0	L	10L/kg

The following procedure illustrates the synthesis of a compound of formula 4:

under a nitrogen atmosphere, 88.0kg of the compound of formula 3, 880.0L of methylene chloride and 11.0kg of dimethylformamide were added to a clean, dry, glass-lined vessel. To the mixture was added 89Kg of thionyl chloride while keeping the temperature not lower than 30 ℃ during the addition. The contents of the reaction vessel were heated at reflux temperature for a minimum of 5 hours before sampling indicated completion of the reaction, and if necessary, the pH was adjusted with 50% NaOH, which was maintained at 7.0-8.0 and the reaction temperature was maintained below 25 ℃. The two-phase mixture was stirred for 15-20 minutes for at least 30 minutes. The phases were separated and the organic layer was concentrated to its volume of 1/3 by removing dichloromethane. 880L of heptane was added and the remainder of the dichloromethane was distilled continuously until the distillate temperature reached 65-68 ℃. The mixture was cooled to 10-15 ℃ over 5 hours, the solid isolated by filtration was granulated for a minimum of 1 hour and washed with 220L heptane. The solid (compound of formula 4) is dried under vacuum at 45-50 ℃. Example 2: alternative preparation of the Compound of formula 4

Sodium bicarbonate can be successfully used instead of sodium hydroxide in the reaction shown in example 1, as shown in this example:

material	Measurement of	Unit of	Equivalent weight/volume
				A compound of formula 3	30.0	kg	1 equivalent of
Thionyl chloride	36.4	kg	3 equivalents of
				Dimethyl formamide	3.75	kg	0.5 equivalent
Methylene dichloride	300	L	10L/kg
				50% sodium hydroxide aqueous solution	According to the need	L
Heptane (Heptane)	375	L	12.5L/kg
				Heptane (Wash)	90	L	3L/kg
Sodium bicarbonate	64.2	kg	7.5 equivalents

Under nitrogen atmosphere, 30.0kg of the compound of formula 3, 300.0L of dichloromethane and 3 were added.75kg of dimethylformamide was added to a clean, dry, glass-lined vessel. To the mixture was added 36.4Kg of thionyl chloride while keeping the temperature not lower than 30 ℃ during the addition. The contents of the reaction vessel were heated at reflux temperature for a minimum of 13 hours before sampling indicated completion of the reaction. The reaction mixture was cooled to 20-25 ℃ andadded to a stirred solution of 64.2kg sodium bicarbonate and 274L water cooled to 4 ℃ to maintain its temperature below 10 ℃. If desired, the pH is adjusted with 50% NaOH, which is maintained at 7.0-8.0. The two-phase mixture was stirred for 15-20 minutes and held at 10-20 ℃ for a minimum of 30 minutes. The phases were separated and the organic layer was concentrated to its volume of 1/3 by removing dichloromethane. 375L of heptane was added and the remainder of the dichloromethane was distilled continuously until the distillate temperature reached 65-68 ℃. The mixture was cooled to 0-5 ℃ over 4 hours, the solid isolated by filtration was granulated for a minimum of 1 hour and washed with 90L heptane. The solid (compound of formula 4) is dried under vacuum at 45-50 ℃. Example 3: preparation of compounds of formulae 6 and 2 (step 2): reaction:

the following materials were used to synthesize the compound of formula 6 and the compound of formula 2 as intermediates:

material	Measurement of	Unit of	Equivalent weight/volume
				A compound of formula 5	61.1	kg	1.2 equivalents
Toluene	489	L	8L/kg (corresponding to the compound of formula 5)
				Sodium hydroxide pellets	4.5	kg	0.16 equivalent
Filter aid	0.5	kg	0.017kg/kg (corresponding to the compound of formula 5)
				Formula 4Compound (I)	90.8	kg	1.0 equivalent
Acetonitrile	732	L	12L/kg (corresponding to the compound of formula 5)

Example 4: preparation of the Compound of formula 2

The following procedure illustrates the synthesis of compounds of formula 2 and intermediates of formula 6:

61.1kg of the compound of formula 5, 4.5kg of sodium hydroxide pellets and 489L of toluene were charged into a clean, dry reaction vessel under a nitrogen atmosphere, and the reaction temperature was adjusted to 105-108 ℃. The acetone was removed by atmospheric distillation over 4 hours while adding toluene to maintain 6L of solvent per kg of compound of formula 5. The reaction mixture was then heated at reflux temperature and the distillate was returned to the tank until the reaction was complete. The mixture was then cooled to 20-25 ℃ at which time a slurry of 40.0L of toluene and 0.5kg of filter aid was added to the reaction mixture and the mixture was stirred for 10-15 minutes. The resulting material was filtered to remove the filter aid, and the filter cake (compound of formula 6) was washed with 30L of toluene.

The filtrate (compound of formula 6) was placed in a clean, dry reaction vessel under nitrogen atmosphere, and 90.8kg of compound of formula 4 and 732L of acetonitrile were added to the reaction vessel. The reaction vessel was heated to reflux temperature with good stirring. The stirring speed was reduced when solids appeared. When the reaction is complete, the contents of the reactor are cooled to 19-25 ℃ over 3-4 hours and the contents are stirred at a temperature of 20-25 ℃ for at least 1 hour. The solid (compound of formula 2, polymorph form a, or a mixture of polymorphsa and B) is then isolated by filtration, and the filter cake is washed with 50L acetonitrile x 2 and dried under vacuum at 45-50 ℃.

It has been found that by reducing the amount of acetonitrile (relative to toluene) the formation of the form a polymorph is favored, and if isopropanol is substituted for acetonitrile, the form a polymorph is favored. However, the use of propanol or other alcohols as co-solvents is not preferred because the oxygen of the alcohol and the 4-carbon of the quinazoline tend to form an ether linkage rather than the desired ethynylphenylamino moiety.

It has further been found that adjusting the pH of the reaction to a pH of 1 to pH7, preferably to a pH of 2 to pH5, more preferably to a pH of 2.5 to pH4, and most preferably to a pH of 3, improves the rate of the reaction. Example 5: the compound of formula 2 (which may be the a form polymorph, or a mixture of the a and B forms polymorphs) is recrystallized to the B form polymorph (step 3): reaction of

The following materials are used to convert the form a polymorph (or a mixture of form a and form B polymorphs) of the compound of formula 2 to the form B polymorph.

Material	Measurement of	Unit of	Equivalent weight/volume
				A type polymorph (formula 2)	117.6	kg	1 equivalent of
2B-ethanol	1881.6	L	16L/kg
				Water (W)	470.4	L	4L/kg

The following procedure illustrates the conversion of the form a polymorph (or a mixture of forms a and B polymorph) of the compound of formula 2 to the thermodynamically more stable form B polymorph:

117.6kg of the form A polymorph (or a mixture of the form A and B polymorphs) was added to a clean, dry reaction vessel under nitrogen atmosphere along with 1881.6L of 2B-ethanol and 470.4L of water. The temperature was adjusted to reflux temperature (about 80 ℃), and the mixture was stirred until the solid dissolved. The solution was cooled to 65-70 ℃ and filtered to be clear. The solution was further cooled to 50-60 ℃ over a minimum of 2 hours with slow stirring, and the precipitate was granulated at this temperature for 2 hours. The mixture is further cooled to 0-5 ℃ in a minimum of 4 hours, at which temperature the precipitate is granulated for 2 hours. The solid (polymorph form B) was isolated by filtration and washed with at least 100L 2B-ethanol. The solid crystalline form was determined to be the form B polymorph of [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride substantially free of the form a polymorph. The solid obtained by this process is a substantially homogeneous polymorph form B (corresponding to the polymorph form a). The process can produce at least 70 wt%, at least 80 wt%, at least 90 wt%, at least 95 wt%, at least 98 wt% of the polymorph form B (corresponding to the polymorph form a). It will be appreciated that the process described herein is illustrative only and does not exclude other variations of the above parameters, and that different particle sizes and yields of the form B polymorph may be produced depending on the desired storage, handling and preparative application of the compound. The solid is dried under vacuum at a temperature below 50 ℃ and the final product is comminuted to provide the form B polymorph in a useable form. Example 6: clinical studies using the stable form B polymorph of [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride:

[6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]The stable form B polymorph of (3-ethynylphenyl) -amine hydrochloride is a potent, selective and orally active inhibitor of epidermal growth factor receptor (EFGR) protein-tyrosine kinase, an oncogene involved in the characteristic abnormal growth of cancer cells. The compound was evaluated in clinical trials on normal healthy volunteers and cancer patients to assess its safety profile and efficacy.Phase I clinical study

Phase I clinical studies of the stable form B polymorph of[6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride were performed in healthy volunteers followed by single doses of 25-200 mg/day or 100-1600 mg/week in cancer patients. For the 150 mg/day dose, the study data did not show stronger than moderate side effects. In the daily dose regimen study, the limiting toxicity at the 200 mg/day dose was diarrhea. The use of loperamide (Imodium. RTM.) effectively controlled the side effects observed at the 150 mg/day dose. The second side effect observed in these studies and the most pronounced toxicity at the 150 mg/day dose was a simple acne-like rash similar to those reported for other EGFR inhibitor drugs. The rash is distributed on the waist, including the face, scalp, neck, arms, chest and back. The rash has a unique histopathology of PMN penetration with slight epidermal hyperproliferation. This is different from hypersensitivity of drugs and does not show the symptoms of known named skin diseases. The rash did not significantly hinder the patient from performing phase II clinics. The stable polymorph form B of [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride showed very good tolerability safety in a total of 290 patients tested in phase I clinics and ongoing phase II clinics. Also, preliminary evidence of efficacy was observed in phase I clinics. For example, of the 28 patients in phase I clinics, 8 patients survived more than 1 year and 12 patients survived for 9-22 months after receiving treatment.

In a phase I clinical study, [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]The stable form B polymorph of (3-ethynylphenyl) -amine hydrochloride in combination with one or more other drugs or treatments, preferably selected from one of the following groups, at a dose of 1-7000 mg/day, preferably 5-2500 mg/day, most preferably 5-200 mg/day: paclitaxel, Gemcitabine, Taxotere, Capcitabine, 5FU, cisplatin, Temozolomide, radiation therapy, and chemoradiation therapy.Phase II and III clinics

Stable polymorph form B of [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride at a dose of 150 mg/day began three phase II single dose studies on refractory non-small cell lung cancer, advanced head and neck cancer, and refractory ovarian cancer.

Evidence of single agent anti-tumor activity of the stable form B polymorph of [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride was observed in patients with advanced cancer of several different tumor types. For example, when administered to patients with advanced head and neck cancer, initial findings indicate that the stable form B polymorph of [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride is a well tolerated oral drug with monotherapeutic activity. Preliminary results indicated that 3 patients had partial responses of interest, while 9 other patients showed evidence of stable disease status. Acneiform rash appeared in approximately 70% of the patients in the first group of the study, which is clearly characteristic of all anti-EGFR inhibitors in clinical trials.

Early data in phase II clinics in 48 patients with refractory non-small cell lung cancer (NSCLC) also showed the effectiveness of using the stable type B polymorph of [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride as a single drug treatment anti-tumor agent for NSCLC. Of the 19 patients evaluated at the beginning of the study, 5 showed partial response to the target, while the other 4 showed signs of stable disease status. Partial responses were observed in patients previously treated with two and three different chemotherapeutic regimens. Thus, it is clear that the stable polymorph form B of [6, 7-bis (2-methoxyethoxy) quinazolin-4-yl]- (3-ethynylphenyl) -amine hydrochloride is a well-tolerated oral drug which is active against non-small cell lung cancer.

Patient eligibility criteria for participation in the single drug study required patients to fail platinum-based chemotherapy and have tumors histopathologically confirmed as EGFR positive. In addition to various secondary endpoints, the primary endpoints of the study were the response rate and time-to-progression of stable disease.

Evidence of anti-tumor activity in ovarian cancer patients can be found in ongoing phase II clinics. Preliminary results indicated that 2 patients had partial responses of interest, while 4 other patients showed evidence of stable disease status. Documented evidence of anti-tumor activity of other EGFR-positive tumor types, including colorectal and renal cell carcinomas, can also be found from phase I studies in cancer patients with multiple tumor types.

Claims (54)

1. A substantially homogeneous crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride designated the form B polymorph that exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14 and 26.91.

2. The polymorph according to claim 1, characterized by an X-ray powder diffraction pattern as shown in figure 3.

3. A crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride designated the form B polymorph that exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14and 26.91, that is substantially free of the form a polymorph.

4. The polymorph according to claim 3, further characterized by an X-ray powder diffraction pattern as shown in figure 3.

5. A composition comprising a substantially homogeneous crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, designated the B polymorph, that exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14 and 26.91.

6. A composition according to claim 5, wherein N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta approximately as follows: 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 6.255 100.0 17.668 2.5 22.982 4.8 27.534 0.9 32.652 1.7 7.860 3.2 18.193 0.7 23.589 2.3 28.148 1.5 33.245 1.7 9.553 3.9 18.749 1.5 23.906 3.0 28.617 4.3 34.719 1.5 11.414 1.5 19.379 1.0 24.459 6.8 29.000 1.4 35.737 0.8 12.483 6.4 20.196 14.4 25.138 10.0 29.797 2.1 36.288 1.0 13.385 9.6 20.734 4.2 25.617 3.7 30.267 0.9 36.809 0.6 14.781 2.1 21.103 14.4 25.908 3.9 30.900 1.6 37.269 1.1 15.720 2.9 21.873 4.7 26.527 2.8 31.475 2.2 37.643 1.4 16.959 5.5 22.452 4.5 26.911 5.6 31.815 2.4 38.114 1.7

7. a composition according to claim 5 wherein the N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride in the form of the form B polymorph is characterized by an X-ray powder diffraction pattern as shown in figure 3.

8. A composition comprising a crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride designated the B polymorph, wherein said polymorph exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at about 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14 and 26.91, and wherein the percentage by weight of the B polymorph relative to the a polymorph is at least 70%.

9. A composition according to claim 8, wherein the form B polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta as follows: 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 6.255 100.0 17.668 2.5 22.982 4.8 27.534 0.9 32.652 1.7 7.860 3.2 18.193 0.7 23.589 2.3 28.148 1.5 33.245 1.7 9.553 3.9 18.749 1.5 23.906 3.0 28.617 4.3 34.719 1.5 11.414 1.5 19.379 1.0 24.459 6.8 29.000 1.4 35.737 0.8 12.483 6.4 20.196 14.4 25.138 10.0 29.797 2.1 36.288 1.0 13.385 9.6 20.734 4.2 25.617 3.7 30.267 0.9 36.809 0.6 14.781 2.1 21.103 14.4 25.908 3.9 30.900 1.6 37.269 1.1 15.720 2.9 21.873 4.7 26.527 2.8 31.475 2.2 37.643 1.4 16.959 5.5 22.452 4.5 26.911 5.6 31.815 2.4 38.114 1.7

10. the composition according to claim 8 wherein the form B polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride is characterized by an X-ray powder diffraction pattern as shown in figure 3.

11. A pharmaceutical composition comprising a therapeutically effective amount of the polymorph of claim 1 and a pharmaceutically acceptable carrier.

12. The pharmaceutical composition according to claim 11, wherein the pharmaceutical composition is suitable for oral administration.

13. The pharmaceutical composition according to claim 12, wherein the pharmaceutical composition is in the form of a tablet.

14. A method of treating a hyperproliferative disorder in a mammal which comprises administering to said mammal a therapeutically effective amount of the polymorph of claim 1.

15. The method according to claim 14, wherein the method is for the treatment of a cancer selected from the group consisting of brain, squamous cell, bladder, gastric, pancreatic, breast, head, neck, oesophageal, prostate, colorectal, lung, kidney, ovarian, gynaecological and thyroid cancer.

16. The method according to claim 14, wherein the method is for the treatment of a cancer selected from non-small cell lung cancer (NSCLC), refractory ovarian cancer, head and neck cancer, colorectal cancer and renal cancer.

17. The method according to claim 14, wherein said therapeutically effective amount is from about 0.001 to about 100mg/kg day.

18. The method according to claim 14, wherein said therapeutically effective amount is from about 1 to about 35 mg/kg/day.

19. The method according to claim 14, wherein the therapeutically effective amount is from about 1 to about 7000 mg/day.

20. The method according to claim 19, wherein said therapeutically effective amount is from about 5 to about 2500 mg/day.

21. The method according to claim 20, wherein said therapeutically effective amount is from about 5 to about 200 mg/day.

22. The method according to claim 21, wherein said therapeutically effective amount is from about 25 to about 200 mg/day.

23. A method for treating a hyperproliferative disorder in a mammal comprising administering to said mammal a therapeutically effective amount of the polymorph of claim 1 in combination with an anti-neoplastic agent selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormonal agents, and anti-androgens.

24. A method of preparing a crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride designated the form B polymorph, wherein said method comprises the step of recrystallizing N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride in a solvent comprising an alcohol.

25. The process according to claim 24, wherein the solvent further comprises water.

26. A method according to claim 24, wherein N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride is prepared by reacting a compound of formula 6

Coupling with a compound of formula 4.

27. The process according to claim 26, wherein the compound of formula 6 is prepared by reacting the compound of formula 5 under heating in a suspension of an alkali metal and a solvent.

28. The process according to claim 26, wherein the preparation of the compound of formula 4 is carried out by chlorinating the compound of formula 3.

29. A process for preparing the form B polymorph of claim 1 comprising the steps of:

a) reacting in a solvent mixture of thionyl chloride, dichloromethane and dimethylformamide to substitute the starting quinazolinamine compound of formula 3 having a hydroxyl group chlorinated,

to provide a compound of formula 4;

b) in situ from the starting material of the compound of formula 5 by reacting the compound of formula 5 under heating in a suspension of an alkali metal and a solvent

Preparing a compound of formula 6;

c) reacting the compound of formula 6 in situ with a compound of formula 4, wherein the compound of formula 6 replaces the chlorine in the compound of formula 4 to provide N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride;

d) recrystallization of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride in alcohol gives the form B polymorph.

30. The process according to claim 29, wherein the substitution chlorination reaction is quenched in the presence of aqueous sodium hydroxide.

31. The process according to claim 29 wherein the substitution chlorination reaction is quenched in the presence of aqueous sodium bicarbonate.

32. The process according to claim 29, wherein the substitution chlorination reaction is quenched in the presence of aqueous potassium hydroxide, aqueous potassium bicarbonate, aqueous potassium carbonate, aqueous sodium carbonate, or a mixture thereof.

33. A process for preparing a crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride designated the form B polymorph by recrystallization, wherein said process comprises the steps of:

e) heating and refluxing alcohol, water and N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride to form a solution;

f) cooling the solution to about 65-70 ℃;

g) allowing the solution to settle; and

h) precipitating the form B polymorph by further cooling the clarified solution.

34. A substantially homogeneous crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride designated the form a polymorph characterized by an X-ray powder diffraction pattern as shown in figure 1.

35. A crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride designated the form a polymorph characterized by an X-ray powder diffraction pattern as shown in figure 1 which is substantially free of the form B polymorph.

36. A composition comprising a substantially homogeneous crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, designated the form a polymorph, wherein said form a polymorph is characterized by the following peaks in the X-ray powder diffraction pattern as shown in figure 1.

37. A composition comprising a substantially homogeneous crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride in the form of the form a polymorph, wherein said form a polymorph is characterized by the following peaks:

polymorph form A

Anode: cu-wavelength 1: 1.54056 wavelength 2: 1.54439(Rel Strength: 0.500)

Range # 1-coupled: 3.000-40.000 stages: 0.040 stage: 1.00

Flat width: 0.300 threshold: 1.0 d(A) I(rel) d(A) I(rel) d(A) I(rel) d(A) I(rel) d(A) I(rel) 15.82794 100.0 6.63179 1.7 4.54453 4.8 3.61674 8.2 2.91238 3.5 14.32371 3.9 5.84901 2.1 4.19685 4.7 3.50393 9.3 2.73148 3.7 11.74376 1.5 5.69971 2.3 4.16411 4.4 3.40200 6.0 2.60193 1.8 11.03408 1.2 5.46922 2.4 3.97273 4.7 3.35174 5.3 2.48243 1.3 10.16026 1.4 5.21396 3.6 3.91344 12.4 3.29005 4.2 2.40227 2.2 8.98039 13.1 4.80569 3.5 3.78223 24.2 3.05178 7.1 2.31297 1.7 7.85825 7.8 4.70077 12.2 3.67845 8.8 2.97750 3.0

Alternatively, a-type polymorph

Anode: cu-wavelength 1: 1.54056 wavelength 2: 1.54439(Rel Strength: 0.500)

Range # 1-coupled: 3.000-40.000 stages: 0.040 stage: 1.00

Flat width: 0.300 threshold: 1.0 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 5.579 100.0 13.340 1.7 19.517 4.8 24.594 8.2 30.673 3.5 6.165 3.9 15.135 2.1 21.152 4.7 25.398 9.3 32.759 3.7 7.522 1.5 15.534 2.3 21.320 4.4 26.173 6.0 34.440 1.8 8.006 1.2 16.193 2.4 22.360 4.7 26.572 5.3 36.154 1.3 8.696 1.4 16.991 3.6 22.703 12.4 27.080 4.2 37.404 2.2 9.841 13.1 18.447 3.5 23.502 24.2 29.240 7.1 38.905 1.7 11.251 7.8 18.862 12.2 24.175 8.8 30.007 3.0

38. A composition comprising a substantially homogeneous crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride in the form of the form B polymorph, wherein said form B polymorph is characterized by the following peaks:

polymorph form B

Anode: cu-wavelength 1: 1.54056 wavelength 2: 1.54439(Rel Strength: 0.500)

Range # 1-coupled: 3.000-40.040 grades: 0.040 stage: 1.00

Flat width: 0.300 threshold: 1.0 d(A) I(rel) d(A) I(rel) d(A) I(rel) d(A) I(rel) d(A) I(rel) 14.11826 100.0 5.01567 2.5 3.86656 4.8 3.23688 0.9 2.74020 1.7 11.23947 3.2 4.87215 0.7 3.76849 2.3 3.16755 1.5 2.69265 1.7 9.25019 3.9 4.72882 1.5 3.71927 3.0 3.11673 4.3 2.58169 1.5 7.74623 1.5 4.57666 1.0 3.63632 6.8 3.07644 1.4 2.51043 0.8 7.08519 6.4 4.39330 14.4 3.53967 10.0 2.99596 2.1 2.47356 1.0 6.60941 9.6 4.28038 4.2 3.47448 3.7 2.95049 0.9 2.43974 0.6 5.98828 2.1 4.20645 14.4 3.43610 3.9 2.89151 1.6 2.41068 1.1 5.63253 2.9 4.06007 4.7 3.35732 2.8 2.83992 2.2 2.38755 1.4 5.22369 5.5 3.95667 4.5 3.31029 5.6 2.81037 2.4 2.35914 1.7

Alternatively, the B-type polymorph

Anode: cu-wavelength 1: 1.54056 wavelength 2: 1.54439(Rel Strength: 0.500)

Range # 1-coupled: 3.000-40.040 grades: 0.040 stage: 1.00

Flat width: 0.300 threshold: 1.0 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 2-Theta I(rel) 6.255 100.0 17.668 2.5 22.982 4.8 27.534 0.9 32.652 1.7 7.860 3.2 18.193 0.7 23.589 2.3 28.148 1.5 33.245 1.7 9.553 3.9 18.749 1.5 23.906 3.0 28.617 4.3 34.719 1.5 11.414 1.5 19.379 1.0 24.459 6.8 29.000 1.4 35.737 0.8 12.483 6.4 20.196 14.4 25.138 10.0 29.797 2.1 36.288 1.0 13.385 9.6 20.734 4.2 25.617 3.7 30.267 0.9 36.809 0.6 14.781 2.1 21.103 14.4 25.908 3.9 30.900 1.6 37.269 1.1 15.720 2.9 21.873 4.7 26.527 2.8 31.475 2.2 37.643 1.4 16.959 5.5 22.452 4.5 26.911 5.6 31.815 2.4 38.114 1.7

39. A prodrug of a compound according to claim 1.

40. A method of inducing differentiation of tumor cells in a tumor comprising contacting said cells with an effective amount of a compound of claim 1 or a composition of claim 3 or 6, thereby differentiating said tumor cells.

41. A method for the treatment of NSCLC (non-small cell lung cancer), pediatric malignancies, cervical and other tumors caused or facilitated by Human Papillomavirus (HPV), melanoma, Barrett's esophagus (pre-malignant syndrome), adrenal and skin cancers and autoimmune diseases, neoplastic skin diseases and atherosclerosis in a mammal, wherein said method comprises administering to said mammal a therapeutically effective amount of a composition comprising at least one N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine and pharmaceutically acceptable salts thereof in anhydrous and hydrated forms.

42. A method according to claim 41, wherein the treatment further comprises palliative or neo-adjuvant/adjuvant monotherapy.

43. The method according to claim 41, wherein the treatment further comprises blocking Epidermal Growth Factor Receptor (EGFR).

44. The method according to claim 41, wherein the method is for the treatment of EGFRvIII expressing tumors.

45. A method according to claim 41, wherein the treatment further comprises a combination with any of chemotherapy and immunotherapy.

46. A method according to claim 41, wherein the treatment further comprises simultaneous treatment with either or both of an anti-EGFR and an anti-EGF antibody.

47. A method according to claim 41, wherein said treatment further comprises administering to said mammal an agent selected from the group consisting of MMPs (matrix-metallo-proteases), VEGFR (vascular endothelial growth factor receptors), farnesyl transferase, CTLA₄(cytotoxic T-lymphocyte antigen 4) and erbB2 inhibitors, MAbs of VEGFr, rhuMAb-VEGF, and,erbB2Mab and avb3 Mab.

48. The method according to claim 41, wherein the pharmaceutical compound is used as a radiosensitizer for cancer treatment, or in combination with anti-hormone therapy.

49. The method according to claim 41, wherein said pharmaceutical compound is used in a radiation therapy regimen to inhibit human tumor growth.

50. A method for chemoprevention of basal cell carcinoma or squamous cell carcinoma or of a human at high risk for said carcinoma in the skin of an area exposed to sunlight, said method comprising administering to said human a therapeutically effective amount of a pharmaceutical composition comprising at least one N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine and the anhydrous and hydrate forms of pharmaceutically acceptable salts thereof.

51. A method of inducing differentiation of tumor cells in a tumor, comprising contacting said cells with an effective amount of at least one N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine compound and anhydrous and hydrate forms of a pharmaceutically acceptable salt thereof.

52. A method of preparing a composition comprising a substantially homogeneous crystalline polymorph of N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride, designated the B polymorph, that exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta at approximately 6.26, 12.48, 13.39, 16.96, 20.20, 21.10, 22.98, 24.46, 25.14 and 26.91, comprising mixing the crystalline polymorph of claim 1 with a carrier.

53. The method according to claim 52, wherein the N- (3-ethynylphenyl) -6, 7-bis (2-methoxyethoxy) -4-quinazolinamine hydrochloride of the form B polymorph is characterized by an X-ray powder diffraction pattern as shown in figure 3.

54. The method according to claim 52, wherein the carrier is a pharmaceutically acceptable carrier.

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